The present invention relates to a novel hydantoin derivative represented by the following formula (I) which shows an inhibitory activity against farnesyl transferase, and thus can be used as an effective therapeutic agent against anti-proliferative diseases such as resfenosis, Rheumatitis arthrititis and particularly cancer: 
or pharmaceutically acceptable salts thereof, in which R1, R2, R3 and R4 are defined as described below.
The present invention also relates to a process for preparation of the compound of formula (I) and to an anticancer composition comprising the compound of formula (I) as an active ingredient.
Mammalian Ras proteins act as molecular switches in the signalling events associated with cell growth and differentiation. The ras proto-oncogene family consists of three members, N-, K-, and H-ras, which code for highly homologous four types of proteins; i.e. H-, N-ras proteins of 189 residues and two isomorphic K-ras-4B and K-ras-4A proteins of 188 and 189 residues, respectively. The chemical basis for the switch mechanism involves cycling of the protein between the inactive (off) guanosine diphosphate (GDP) bound state and the active (on) guanosine triphosphate (GTP) bound state (Bourne. H. R.; Sanders, D. A.; McCormick, F.; Nature, 1991, 349,117). Biochemical and structural studies have shown that point mutations of the residues 12, 13, and 61, positioned in the neighborhood of phosphoryl group of GTP, resulting in the decrease of guanosine triphosphatase activity are associated with many human cancers, particularly, pancreatic cancer, urinary bladder carcinoma, colon cancer, etc. (Bos, J. L., Cancel Res., 1989, 49, 4682).
Ras protein is synthesized as a cytosolic precursor that is ultimately localized to the cytoplasmic face of the plasma membrane after a series of posttranslational modification (Gibbs, J. B., Cell 1991, 65, 1). These series of biochemical modifications, by changing the electrical charge state or spacial structure to increase the hydrophobicity allow Ras protein to attach to cell membrane more easily. The first and obligatory step in the series is the addition of a farnesyl moiety to the cysteine residue of the C-terminal CAAX motif (C, cysteine, A, usually aliphatic residue; X, any other amino acid) in a reaction catalyzed by farnesyl protein transferase (FTase). This modifications is essential for Ras function, as demonstrated by the inability of Ras mutants lacking the C-terminal cysteine to be farnesylated, to localize to the plasma, and to transform mammalian cells in culture (Hancock, J. F., Magee, A. I., Child, J. E., Marshall, C. J., Cell 1989, 57, 1167). The subsequent posttranslational modifications, cleavage of the AAX residues, carboxyl methylation of the farnesylated cysteine, and palmitoylation of the cysteines located upstream of the CAAX motif in H-and N-ras proteins are not obligatory for Ras membrane association or cellular transforming activity. Interestingly, K-ras-4B, different from H- and N-ras, has a multiple lysine rich region named polybasic domain, instead of having cysteine required for palmitoylation, thereby facilitating the farnesylated ras protein to bind to anionic lipid layer of cell membrane. The inhibitors of FTase that catalyzes the obligatory modification have therefore been suggested as anticancer agents for tumors in which Ras oncogene contributes to transformation (Buses, J. E. et al., Chemistry and Biology, 1995, 2 787). A number of FTase inhibitors have recently identified demonstrated potent and specific ability to block Ras farnesylation. signalling and transformation in transformed cells and tumor cell lines both in vitro and in animal models (Koh, N. E. et al., Proc. Natl. Acad. Sci. USA, 1994, 91, 914; Kohl, N. E. et al., Nature Medicine. 1995, 1 792). However, most of the inhibitors are related to CAAX motif as Ras substrate mimic and peptidie in nature or contain a sulfhydryl group (U.S. Pat. No. 5.141,851; Kohl, N. E. et al., Science, 1993, 260, 1934; PCT/US95/12224, Graham et al., Sebti, S. M. et al., J. Biol. Chem., 1995, 270, 26802; James, G. L. et al., Science, 1993 260, 1937; Bishop, W. R. et al., J. Biol. Chem., 1995, 270, 3061 1) Recently, a new type of peptidomimetic inhibitor imitating catalytic step of FTase has been reported (Poulter, C. D. et al., J. Am. Chem. Soc., 1996, 118,8761). The chemical basis of the inhibitor design relates to the reaction mechanism. That is, transferring prenyl group by the enzyme is electrophilic displacement and the reaction requires (+) charge in a transition state.
These inhibitors previously described however possess limited activity and selectivity for inhibition of the oncogenic function of Ras proteins, particularly K-ras-4B, which is found to be most common in human cancer. Therefore, new inhibitor having the ability of effectively inhibiting K-ras activity is required.
The present inventors have performed studies for developing a compound having the structure characteristics imitating transition state of catalytic reaction of farnesyl transferase and as a result, found that hydantoin derivatives according to the present invention can inhibit farnesyl transferase activity by imitating transition state of catalytic reaction of farnesyl transferase.
Therefore, the object of the present invention is to provide a hydantoin derivative of formula (I) which inhibits the activity of farnesyl transferase, process for preparation thereof, and anti-cancer composition comprising the compound of formula (I) as an active component.
It is the first object of the present invention to provide a hydantoin derivative represented by the following formula (I) and pharmaceutically acceptable salt thereof which inhibit the activity of farnesyl transferase 
in which
R1 and R2 independently of one another represent hydrogen; lower alkyl; monocyclic or bicyclic alkyl group which can be substituted by lower alkyl or halogen; heterocyclic group containing hetero atoms selected from a group consisting of nitrogen and sulfur as ring member; or a radical having the following formula: 
(wherein D represents alkoxy; hydroxy; amino acid residue; morpholine; thiomorpholine; piperazine, alkoxyalkylamine or alkyloxyalkylamine each of which is substituted or unsubstituted by lower alkyl, and m is selected from 0 to 2),
R3 represents amino acid residue, or a radical having the following formula, 
wherein
A represents hydrogen; lower alkyl; aryl group which is substituted by substituents selected from a group consisting of halogen, cyano (CN) nitro(NO2), carboxy(COOH), amide, thioamide, SR and lower alkyl; heterocyclic group which is substituted by substituents selected from a group consisting, of halogen, cyano, nitro, COOR, amide, thioamide, SR and lower alkyl and which comprises nitrogen or sulfur atom as ring member; lower alkyl substituted by the substituted aryl or heterocyclic group as mentioned above; or a radical having the following formula: 
(in the definition for the substituent A, R represents hydrogen or lower alkyl, and E represents hydrogen or xe2x80x94Fxe2x80x94G wherein F represents CH2, Cxe2x95x90O, SO2, and G represents hydrogen; lower alkyl substituted or unsubstituted by phenyl or biphenyl: lower alkoxy; phenyl benzyl; benzyloxy; amine substituted or unsubstituted by lower alkyl, phenyl, benzyl, cycloalkyl or phenoxy alkyl),
B and C independently of one another represent hydrogen, halogen or lower alkyl.
n denotes an integer of 0 to 4,
R4 represents hydrogen; aromatic group substituted or unsubstituted by lower alkyl or halogen; bicyclic aromatic group; heteroaromatic group containing hetero atoms selected from a group consisting of nitrogen and sulfur as ring member; or a radical having the following formula: 
wherein
R5 represents aryl group substituted by lower alkoxy; or heterocyclic group containing hetero atoms selected from a group consisting of nitrogen, oxygen and sulfur as ring member,
R6 represents hydrogen; lower alkyl; lower alkyl which is substituted by substituents selected from a group consisting of halogen, cyano, hydroxy, COOR, amide, thioamide, SR and SO2R; lower alkyl substituted by an aryl group which is substituted by substituents selected from a group consisting of halogen, cyano, COOR, amide, thioamide, SR, SO2R and lower alkyl; heterocyclic group containing hetero atoms selected from a group consisting of nitrogen and sulfur as ring member; heterocyclic group which is substituted by substituents selected from a group consisting of halogen, cyano, COOR, amide, thioamide, SR, SO2R and lower alkyl and which contains hetero atoms selected from a group consisting of nitrogen and sulfur as ring member, wherein R represents lower alkyl,
R7 and R8 independently of one another represent hydrogen, halogen, halogenoalkyl, cyano, amide, thioamide, alkoxy or phenoxy, or represent a radical having the following formula, 
wherein
Z represents CH2, CO, O, S, SO2, NR9, NHSO2 or NHCOO,
R10 represents hydrogen, lower alkyl, halogenoalkyl, alkoxy, hydroxy, benzyloxycarbonyl or benzyl,
R9 represents hydrogen or lower alkyl, or lower alkyl substituted by aromatic group,
X represents CH2, CO, O, S or SO2, and
n denotes an integer of 0 to 4.
In the definitions for the substituents of the compound of formula (I), the term xe2x80x9clower alkylxe2x80x9d means a straight or branched alkyl having 1 to 4 carbon atoms which includes methyl, ethyl, isopropyl, isobutyl and t-butyl; the term xe2x80x9cheterocyclic group containing hetero atoms selected from a group consisting of nitrogen and sulfur as ring memberxe2x80x9d means mono- or bicyclic aliphatic or aromatic group containing one or two nitrogen or sulfur in the ring as ring member.
The abbreviations for amino acids used in the present specification are consistent with IUPAC-IUB Commission on biochemical nomenclature of amino acid and peptide[Eur. J. Biochem., 1984, 158, 9-31].
Also, the compound of formula (I) according to the present invention can form a pharmaceutically acceptable salt. Such salt includes non-toxic acid addition salt containing pharmaceutically acceptable anion, for example a salt with inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydriodic acid, etc., a salt with organic carboxylic acids such as tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trofluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, etc., or a salt with sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, etc.; and metal addition salt for example a salt with alkali metal or alkaline earth metal such as lithium salt. Further, the present invention includes a solvate of the compound of formula (I) such as alcoholate, and hydrate thereof.
Since the compound of formula (I) according to the present invention may have asymmetric carbon atoms depending on the substituents, they can be present in the form of racemate, diastereomer mixture or the individual diastereomers. Thus, the present invention also includes all of these stereoisomers and their mixtures.
Among the compound of formula (I) according to the present invention, the preferred compounds include those wherein R1 represents hydrogen; monocyclic or bicyclic aryl group which can be substituted by lower alkyl or halogen; or a radical having the following formula: 
(wherein D represents alkoxy; hydroxy; amino acid residue; morpholine; thiomorpholine; piperazine or alkoxyalkylamine each of which is substituted or unsubstituted by lower alkyl, and m is selected from 0 to 1),
R2 represents hydrogen; lower alkyl; or a radical having the following formula: 
wherein D represents alkoxy; hydroxy; amino acid residue; morpholine; thiomorpholine; piperazine or alkoxyalkylamine each of which is substituted or unsubstituted by lower alkyl, and m is selected from 0 to 1),
R3 represents a radical having the following formula, 
wherein
A represents hydrogen; aryl group which is substituted by substituents selected from a group consisting of halogen, cyano(CN), nitro(NO2), carboxy(COOH), amide, thioamide, SR and lower alkyl; or a radical having the following formula: 
(in the definition for the substituent A, R represents hydrogen or lower alkyl, and E represents hydrogen or xe2x80x94Fxe2x80x94G wherein F represents Cxe2x95x90O. and
G represents benzyloxy, lower alkoxy, or lower alkyl substituted or unsubstituted by phenyl),
B and C independently of one another represent hydrogen,
n denotes an integer of 1 to 3,
R4 represents hydrogen; aromatic group substituted or unsubstituted by halogen; bicyclic aromatic group; heteroaromatic group containing hetero atoms selected from a group consisting of nitrogen and sulfur as ring member; or a radical having the following formula: 
wherein
R5 represents aryl group substituted by lower alkoxy; or heterocyclic group containing hetero atoms selected from a group consisting of nitrogen, oxygen and sulfur as ring, member,
R6 represents hydrogen; lower alkyl; lower alkyl which is substituted by substituents selected from a group consisting of halogen, cyano, hydroxy, COOR, amide, thioamide, SR and SO2R; lower alkyl substituted by an aryl group which is substituted by substituents selected from a group consisting of halogen, cyano, COOR, amide, thioamide, SR, SO2R and lower alkyl; or heterocyclic group containing hetero atoms selected from a group consisting, of nitrogen and sulfur as ring member; heterocyclic group which is substituted by substituents selected from a group consisting of halogen, cyano, COOR, amide, thioamide, SR, SO2R and lower alkyl and which contains hetero atoms selected from a group consisting of nitrogen and sulfur as ring member, wherein R represents lower alkyl,
R7 and R8 independently of one another represent hydrogen, halogen, halogenoalkyl, cyano or phenoxy, or represent a radical having the following formula, 
wherein
Z represents O, S, SO2, NR9, NHSO2 or NHCOO,
R10 represents hydrogen, lower alkyl, halogenoalkyl, alkoxy, hydroxy or benzyloxycarbonyl,
R9 represents or lower alkyl,
X represents O, S or SO2, and
n denotes an integer of 1 or 3.
Typical examples of the compound of formula (I) according to the present invention are presented in the following Tables 1a to 1v.
It is another object of the present invention to provide a process for preparing the hydantoin derivative of formula (I) as defined above.
According to the present invention, the hydantoin derivative of formula (I) can be prepared by a process characterized in that
1) a compound represented by the following formula (II): 
wherein R1, R2 and R4 are defined as previously described, is reacted under Mitsunobu reaction condition with an alcohol derivative represented by the following formula (III):
R3xe2x80x94OHxe2x80x83xe2x80x83[Formula III]
wherein R3 is defined as previously described, or
2) a compound represented by the following formula (IIa): 
wherein R1 and R2 are defined as previously described, is reacted with the alcohol derivative of formula (III) under Mitsunobu reaction condition to produce a compound represented by the following formula (Ia): 
wherein R1, R2 and R3 are defined as previously described, then substituent R4xe2x80x2 is introduced into the resulting compound of formula (Ia) to produce a compound represented by the following formula (Ib): 
wherein R1, R2 and R3 are defined as previously described and R4xe2x80x2 is the same as R4 except that R4xe2x80x2 is not hydrogen.
While, the starting compounds used in the above reaction may be prepared according to the processes depicted in the following Schemes 1 to 9. First, the compound of formula (IIa) can be synthesized by condensing a ketone compound with potassium cyanide and ammonium carbonate as represented in the following Reaction Scheme 1: 
As represented in the following Reaction Scheme 2, the compound of formula (II) can be prepared from ethyl bromoacetate by alkylation of amine compound and cyclization: 
Further, the compound of formula (II) may be obtained by processes represented in the following Reaction Schemes 3 and 4. Specifically, a racemate can be obtained through the alkylation of an amine compound using fumarate and then cyclization as Reaction Scheme 3, and a stereospecific isomer can be obtained through the reductive amination of an aspartic acid having ester protecting group with an aldehyde compound and then cyclization as Reaction Scheme 4: 
The alcohol compound of formula (III) may be prepared by processes described in the following Reaction Schemes 5 to 9. That is, the compound 3 can be synthesized by forming imidazole moiety using dihydroxy acetone as Reaction Schemes 5 and 6, or by alkylating the existing imidazole methanol compound as Reaction Scheme 7. Also, the compound of formula (III) may be prepared by processes described in the following Reaction Schemes 8 and 9. 
In the above Reaction Schemes, AcOH represents acetic acid, Cbz-Cl represents benzyloxycarbonyl chloride, Trt-Cl represents chlorotriphenylmethane and TFA represents trifluoroacetic acid. The process for preparing the compound of formula (I), particularly the synthetic methods as described above will be more specifically explained by the following Preparations and Examples.
As the coupling agent used in the above amidation reaction for preparing the compound of formula (I), a mixture of carbodiimides such as dicyclohexylcarbodiimide(DCC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide(EDC), 1,1xe2x80x2-dicarbonyldiimidazole(CDI), etc., and 1-hydroxybenzotrizole can be mentioned.
Reaction conditions including the amount of reactants, temperature, reaction time, etc. can be easily determined by the person having ordinary skill in the art depending on the reactant used.
Also, the free compound of formula (I) produced in the aforementioned reaction may be easily converted into a salt form thereof according to the conventionally known methods in this art.
After the reaction is completed, the resulting product may be further separated and purified by usual work-up processes, such as for example, chromatography, recrystallization, etc.
The compound of formula (I) shows an inhibitory, activity against farnesyl transferase, and thus can be effectively used as an anti-cancer agent.
Therefore, the present invention also provides an anti-cancer agent comprising the novel compound of formula (I), as defined above, or a pharmaceutically acceptable salt thereof as an active ingredient together with a pharmaceutically acceptable carrier.
When the active compound according to the present invention is used for clinical purpose, it is preferably administered in an amount ranging from 10 mg to 40 mg per kg of body weight a day. The total daily dosage may be administered in one time or over several times. However, the specific administration dosage for the patient can be varied with the specific compound used, body weight of the subject patient, sex, hygienic condition, diet, time or method of administration, excretion rate, mixing ratio of the agent, severity of the disease, to be treated, etc.
The compound of the present invention may be administered in the form of injections or oral preparations. Injections, for example, sterilized aqueous or oily suspension for injection, can be prepared according to the known procedure using suitable dispersing agent, wetting agent, or suspending agent. Solvents which can be used for preparing injections include water, Ringer""s fluid and NaCl solution, and also sterilized fixing oil may be conveniently used as the solvent or suspending media. Any non-stimulative fixing oil including mono-, di-glyceride may be used for this purpose. Fatty acid such as oleic acid may also be used for injections.
As the solid preparation for oral administration, capsules, tablets, pills, powders and granules, etc., preferably capsules and tablets can be mentioned. It is also desirable for tablets and pills to be formulated into enteric-coated preparation. The solid preparations may be prepared by mixing the active compound of formula (I) according to the present invention with at least one carrier selected from a group consisting of inactive diluents such as sucrose, lactose, starch, etc., lubricants such as magnesium stearate, disintegrating agent and binding agent.
The present invention will be more specifically explained in the following examples. However, it should be understood that the following examples are intended to illustrate the present invention but not in any manner to limit the scope of the present invention. Processes for preparing the starting substances used for obtaining the compound of formula (I) will be explained in the following Preparations.
30 g(0.18 mol) of 1xe2x80x2-acetonaphthone and 23 g(0.35 mol) of potassium cyanide[KCN] were dissolved in 900 ml of methanol. Then, 169 g(1.8 mol) of ammonium carbonate[(NH4)2CO3] in 900 ml of distilled water were added thereto and the resulting solution was stirred for 12 hours at 70xc2x0 C. The reaction solution was distilled under reduced pressure to remove methanol and the residue was extracted with ethyl acetate(500 mlxc3x974). The extract was distilled under reduced pressure to remove ethyl acetate and 38.2 g (Yield: 90%) of title compound was obtained.
1H NMR(CDCl3) xcex4(ppm) 2.15(3H, s), 6.35(1H, s), 7.44(1H, t), 7.53(2H, m), 7.70(1H, d), 7.89(1H, d), 7.93(1H, d), 7.99(1H, d), 8.52(1H, br)
FAB Mass(M+H): 241
19.3 g(139 mmol) of potassium carbonate was added to 200 ml of dimethylformamide, and then the solution was heated to dissolve potassium carbonate. After cooling to room temperature, 8 ml(70 mmol) of ethyl bromoacetate and 10 g(70 mmol) of 1-naphthylamine were added to the solution, which was then stirred for 48 hours. DMF was removed under reduced pressure and ethyl acetate was added to the residue. The ethyl acetate layer was washed with water 4 times and saturated sodium chloride solution. Ethyl acetate was removed under reduced pressure, and then column chromatography was performed using a mixed solution(9:1) of hexane and ethyl acetate as an eluent to obtain 12 g of the title compound(Yield 75%, MW 229).
1H NMR(CDCl3) xcex4(ppm) 1.33(3H, t), 4.07(2H, s), 4.30(2H, q), 6.24(1H, d), 7.30(1H, d), 7.34(1H, t), 7.47(2H, m), 7.80(1H, m), 7.95(1H, m)
FAB (M+H) 230 2-2) Synthesis of 1-naphthalen-1-yl-imidazolidin-2,4-dione
8.68 g(37.9 mmol) of the compound prepared in Preparation 2-1) and 6.34 g(75.8 mmol) of potassium isocyanate were added to 100 ml of acetic acid and the solution was stirred for 24 hours at 110xc2x0 C. Acetic acid was removed under reduced pressure and then ethyl acetate was added to the residue. The resulting, solution was washed with water 3 times, 1N HCl solution, saturated sodium bicarbonate solution, and saturated sodium chloride solution, respectively. The solution was recrystallized from ethyl acetate to obtain 6.8 g of the title compound(Yield 80%, MW 226).
1H NMR(CD3OD) xcex4(ppm) 4.49(2H, s), 7.52-7.61(4H, m), 7.89-7.98(3H, m)
FAB (M+H) 227
3.12 ml(19.0 mmol) of diethyl fumarate and 3.0 g(19 mmol) of 1-aminomethyl naphthalene were added to acetonitrile, and then the solution was refluxed for 12 hours. Acetonitrile was removed under reduced pressure and column chromatography was performed using a mixed solution of hexane and ethyl acetate(3:1) as an eluent to obtain 4.57 g of the title compound(Yield 73%, MW 329).
1H NMR (CDCl3) xcex4(ppm) 1.19(3H, t), 1.30(3H , t), 2.18(1H, br), 2.70 (2H, 2dd), 3.79(1H, dd), 4.03-4.13(2H, m) 4.15(1H, d), 4.23(2H, q), 4.35 (1H, d), 7.41(1H t), 7.45-7.55(3H, m), 7.77(1H, d), 7.85(1H, d), 8.20(1H, d)
FAB (M+H) 330
4.57 g(13.9 mmol) of the compound prepared in Preparation 3-1) and 3.38 g(41.7 mmol) of potassium isocyanate were added to 150 ml of acetic acid and the solution was stirred for 24 hours at 110xc2x0 C. Acetic acid was removed under reduced pressure and ethyl acetate was added to the residue. The resulting solution was washed with water 3 times, 1N HCl solution, saturated sodium bicarbonate solution and saturated sodium chloride solution, respectively. Ethyl acetate was removed under reduced pressure and then column chromatography was performed using a mixed solution of hexane and ethyl acetate(1:1) as an eluent to obtain 3.85 g of the title compound(Yield 85%, MW 326).
1H NMR (CDCl3) xcex4(ppm) 1.08(3H, t), 2.70(2H, 2dd), 3.80(1H, m), 3.90(2H, m), 4.80(1H, d), 5.30(1H, d), 7.38-7.46(2H, m), 7.51(1H, t), 7.57(1H, m), 7.83(1H, m), 7.87(1H, d), 8.10(1H, d), 9.27(1H, s)
FAB (M+H) 327
2.22 g(11.2 mmol) of (S)-dimethyl aspartate hydrochloride and 1.6 ml(11.2 mmol) of 1-naphthyl aldehyde were added to 50 ml of dimethylformamide, and then the solution was stirred for 1 hour. 5.0 g(22.4 mmol) of sodium triacetoxy borohydride was added to the solution and the resulting solution was stirred for 4 hours. Then, DMF was removed in vacuo and then ethyl acetate was added thereto. The solution was washed with water and saturated sodium chloride solution. Ethyl acetate was removed under reduced pressure and column chromatography was performed on the residue using a mixed solution of hexane and ethylacetate(3:1) as an eluent to obtain 3.00 g of the title compound(Yield 89%, MW 301).
1H NMR (CDCl3) xcex4(ppm) 2.23(1H, br), 2.74(2H, 2dd), 3.60(3H, s), 3.77 (3H, s), 3.81(1H, dd), 4.15(1H, d), 4.34(1H, d), 7.41(1H, t), 7.44-7.55(3H, m), 7.77(1H, d), 7.84(11H d), 8.18(1H, d)
FAB (M+H) 302
3.00 g(9.96 mmol) of the compound prepared in Preparation 4-1) and 2.2 g (26 mmol) of potassium isocyanate were added to 50 ml of acetic acid and then the solution was stirred for 30 minutes at 110xc2x0 C. Acetic acid was removed under reduced pressure and ethyl acetate was added to the residue. The resulting solution was washed with water 3 times, 1N HCl solution, saturated sodium bicarbonate solution and saturated sodium hydroxide solution, respectively. Ethyl acetate was removed under reduced pressure to obtain 2.87 g of the title compound(Yield 92%, MW 312).
1H NMR (CD3OD+CDCl3) xcex4(ppm) 2.45(2H, 2dd), 3.09(3H, s), 3.75(1H, t), 4.79(2H, dd), 7.20(1H, m), 7.27-7.37(2H, m), 7.63(1H, d), 7.66(1H, d), 7.90 (1H, d)
FAB (M+H) 313
22.2 g(0.2 mol) of 4-aminomethyl piperidine was dissolved in 250 ml of toluene and 21.2 g(0.2 mol) of benzaldehyde was added thereto. The reaction mixture was heated to reflux for 3 hours with Dean-stack and then cooled down to 0xc2x0 C. 34.2 g(0.2 mol) of benzyl chloroformate was added dropwise while stirring. The reactants were stirred for 3 hours and 220 ml of 1N KHSO4 was added at room temperature. The reaction solution was extracted with 200 ml of diethylether 3 times and the aqueous layer was basified with sodium hydroxide. After the aqueous solution was treated with saturated sodium chloride solution, it was extracted with 100 ml of dichloromethane 3 times. and the organic layer was dried over magnesium sulfate. Removal of dichloromethane under reduced pressure provided 38 g of the title compound(Yield 91%, MW 248).
1H NMR(CDCl3) xcex4(ppm) 1.11(2H, s), 1.49(3H, s), 1.70(2H, d), 2.57(2H, d), 2.78(2H, s), 4.20(2H, s), 5.12(2H, s), 7.34-7.35(5H, m)
FAB (M+H) 249
24.8 g(0.1 mol) of the compound prepared in Preparation 5-1) was dissolved in 50 ml of n-butanol with 6.0 g(0.1 mol) of acetic acid. Above solution was added to 50 ml of n-butanol solution in which 12.6 g(0.13 mol) of potassium thiocyanate, 15.2 g(0.1 mol) of 1,3-dihydroxyacetone dimer and 10.0 g(0.17 mol) of acetic acid were dissolved, and then the resulting solution was stirred for 48 hours. After stirring, the solvent was removed under the reduced pressure and 200 ml of ethyl acetate was added thereto. The resulting solution was washed with 100 ml of water 3 times and the organic layer was dried over magnesium sulfate. The solvent was removed under reduced pressure to obtain 27 g of the title compound(Yield 75%, MW 361).
1H NMR(CDCl3) xcex4(ppm) 1.22(2H, d), 1.57(2H, d), 2.30(1H, s), 2.72(2H, s), 3.96(2H, s), 4.15(2H, d), 4.46(2H, s), 5.10(2H, s), 6.62(1H, s), 7.26-7.37(5H, m)
FAB (M+H) 362
18.05 g(50 mmol) of the compound prepared in Preparation 5-2) was added to a mixed solution of 100 ml of 10% nitric acid solution and 10 ml of ethyl acetate at 0xc2x0 C. It was stirred for 3 hours at room temperature. The reaction solution was basified with 4N aqueous sodium hydroxide solution and then extracted with 100 ml of ethyl acetate twice. The extracted organic solution was dried over magnesium sulfate. The solvent was removed under reduced pressure to give 12.3 g of the title compound(Yield 75%, MW 329).
1H NMR(CDCl3) xcex4(ppm) 1.16(2H, d), 1.56(2H, d), 1.98(1H, s), 2.70(2H, s), 3.88(2H, d), 4.18(2H, s), 4.49(1H, s), 4.56(3H, s), 5.10(2H, s), 6.82 (1H, s), 7.27-7.40(5H, m)
FAB (M+H) 330
8.9 g(40 mmol) of 4-bromobenzylamine hydrochloride and 4 ml of acetic acid were dissolved in 85 ml of n-butanol. 5.19 g(50 mmol) of potassium thiocyanate and 3.21 g(20 mmol) of 1,3-dihydroxyacetone dimer were added to the solution. The resulting solution was stirred for 4 days and then filtered under reduced pressure to separate the precipitated solid, which was then washed with water and diethylether. The solid thus obtained was added to 10% nitric acid solution, and then the mixture was stirred for 3 hours, filtered under reduced pressure to remove the insoluble impurities. Then, the solution was basified with 4N sodium hydroxide solution to precipitate a solid product. This solid product was washed with water several times and dried under vacuum to give 6.7 g of the title compound(Yield 60%, MW 266).
1H NMR(CDCl3) xcex4(ppm) 4.45(2H, s), 5.20(2H, s), 6.94(1H, s), 7.03(2H, d), 7.18(4H, m)
FAB Mass (M+H) 267
7.98 g(59.2 mmol) of hydroxymethyl imidazole hydrochloride was dissolved in a solvent mixture of 60 ml of dimethylformamide and 20 ml of triethylamine. 200 ml of dimethylformamide solution containing 18.7 g (67 mmol) of triphenylmethyl chloride was added slowly thereto. After 2 hours, 1000 ml of ice water was added thereto to obtain a solid. This solid was recrystallized from dioxane to give 17.6 g of the title compound(Yield 87%, MW 340).
mp 227-229xc2x0 C.
10.0 g(29.4 mmol) of the compound prepared in Preparation 7-1) was added to 200 ml of pyridine and then, 3.30 g((32.4 mmol) of acetic anhydride was added. After stirring for 24 hous at room temperature pyridine was removed under reduced pressure. The residue was dissolved in 400 ml of ethylacetate, and then washed with 200 ml of saturated sodium chloride solution. After removal of ethyl acetate under reduced pressure, chromatography was performed on the residue using dichloromethane/methanol(95:5) as an eluent to obtain 10.44 g of the title compound(Yield 93%, MW 382).
1H NMR(CDCl3) xcex4(ppm) 2.01(3H, s), 4.95(2H, s), 6.88(1H, s), 7.08(5H, s), 7.27(10H, s), 7.45(1H, s)
FAB (M+H) 383
10.0 g(26.2 mmol) of the compound prepared in Preparation 7-2) was dissolved in 40 ml of dichloromethane and 5.64 g(28.8 mmol) of 4-cyanobenzyl bromide was added thereto. The resulting solution was stirred for 60 hours at room temperature and dichloromethane was removed under reduced pressure. Column chromatography was performed with the residue using dichloromethane/methanol(95:5) as an eluent to obtain 10.62 g of the title compound(Yield 70%, MW 578).
1H NMR(CDCl3/CD3OD) xcex4(ppm) 1.95(3H, s), 4.95(2H, s), 5.45(2H, s), 7.11-7.40(18H, m), 7.65(2H, d), 8.21(1H, s)
FAB (M+H) 579
9.10 g(15.7 mmol) of the compound prepared in Preparation 7-3) was dissolved in 500 ml of dichloromethane and 6.06 ml (78.7 mmol) of trifluoroacetic acid and 12.5 ml (78.7 mmol) of triethylsilane were added slowly thereto at 0xc2x0 C. The resulting solution was stirred for 1 hour at room temperature. After removal of dichloromethane under reduced pressure, pH was adjusted to pH 10 using saturated potassium carbonate solution. This solution was extracted with 300 ml of ethyl acetate and ethyl acetate was removed under reduced pressure. Column chromatography was performed with the residue using ethyl acetate as an eluent to obtain 3.60 g of the title compound(Yield 90%, MW 255).
1H NMR(CDCl3) xcex4(ppm) 1.90(3H, s), 4.97(2H, s), 5.25(2H, s), 7.14(2H, d), 7.21(1H, d), 7.67(1H, s), 7.75(2H, d)
FAB (M+H) 256
3.36 g(13.2 mmol) of the compound prepared in Preparation 7-4) was dissolved in 160 ml of methanol and 3.60 g(26.3 mmol) of K2CO3 was added thereto. After stirring for 20 minutes at room temperature, methanol was removed under reduced pressure and product was extracted with 250 ml of ethyl acetate. Column chromatography was performed using dichloromethane/methanol(95:5) as an eluent to obtain 2.55 g, of the title compound(Yield 91%, 0%, MW 213).
1H NMR (CDCl3/CD3OD) xcex4(ppm) 4.28(2H, s), 5.18(2H, s), 6.84(1H, s), 7.12(2H, d), 7.42(1H, s), 7.55(2H, d)
FAB (M+H) 213
5.0 g(73.4 mmol) of imidazole and 12.6 g(148.6 mmol) of methyl acrylate were dissolved in 100 ml of acetonitrile, and then refluxed for 8 hours. Acetonitrile and the excess methyl acrylate was removed under reduced pressure. Then, 200 ml of ethyl acetate was added to the residue and the solution thus obtained was washed with saturated sodium chloride solution. Removal of ethyl acetate under reduced pressure provided 11.1 g(Yield: 90%) of the title compound.
1H NMR(CDCl3) xcex4(ppm) 2.75(2H, t), 3.46(3H, s), 4.24(2H, t), 6.89(1H, s), 7.00(1H, s), 7.46(1H, s)
FAB Mass(M+H): 169
To the 1.1 g(6.6 mmol) of the compound prepared in Preparation 8-1) in 50 ml of tetrahydrofuran was added 0.26 g(6.6 mmol) of lithium aluminum hydride[LiAlH4] and then the resulting mixture was refluxed for one hour. Then, 20 ml of 1N sodium hydroxide solution was added to the reaction mixture which was then extracted with ethyl acetate. Removal of the organic solvent under reduced pressure provided 0.77 g(Yield: 93%) of the title compound was obtained.
1H NMR(CDCl3) xcex4(ppm) 1.67(2H, m), 3.26(2H, t), 3.78(2H, t), 6.60(1H, s), 6.75(1H, s), 7.14(1H, s)
FAB Mass(M+H): 127
5.0 g (73.4 mmol) of imidazole and 3.36 ml(29.4 mmol) of ethyl bromoacetate were dissolved in 50 ml of dimethylformamide and stirred for 4 hours. Dimethylformamide was removed in vacuo. Then, 100 ml of ethyl acetate was added to the residue and it was washed with saturated sodium chloride solution. Removal of the organic solvent under reduced pressure provided 0.77 g(Yield: 17%) of the title compound.
1H NMR(CDCl3) xcex4(ppm) 1.29(3H, t), 4.25(2H, q), 4.70(2H, s), 6.95(1H, s), 7.10(1H, s), 7.49(1H, s)
FAB Mass(M+H): 155
To the 20 ml of tetrahydrofuran solution of 0.77 g(5.0 mmol) of the compound prepared in Preparation 9-1) was added 0.2 g(5.0 mmol) of lithium aluminum hydride and then the resulting mixture was refluxed for one hour. Then, 10 ml of 1N sodium hydroxide solution was added to the reaction mixture, and it was extracted with ethyl acetate. The organic solvent was removed under reduced pressure and the residue was subjected to chromatography using a solvent mixture of methanol-methylene chloride(5:95) as the eluent to obtain 0.51 g(Yield: 91%) of the title compound.
1NMR(CDCl3) xcex4(ppm) 3.78(2H, t), 3.98(2H, t), 6.85(1H, s), 6.94(1H, s), 7.3(1H, s)
FAB Mass(M+H): 113